Simple and highly sensitive molecular diagnosis of Zika virus by lateral flow assays

Dohwan Lee, Yong Shin, Seok Chung, Kyo Seon Hwang, Dae Sung Yoon, Jeong Hoon Lee

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30 Citations (Scopus)

Abstract

We have developed a simple, user-friendly, and highly sensitive Zika virus (ZIKV) detection method by incorporating optimized reverse transcription loop-mediated isothermal amplification (RT-LAMP) and a lateral flow assay (LFA). The optimized RT-LAMP reaction was carried out using Bst 3.0 polymerase, which has robust and fast isothermal amplification performance even in the presence of high concentrations of inhibitors; this permitted the amplification of ZIKV RNA in pure water and human whole blood. In addition, the strong reverse transcription activity of Bst 3.0 polymerase enabled specific ZIKV RNA amplification without extra addition of reverse transcriptase. The RT-LAMP condition was optimized by adjusting the Mg2+ and dNTP mix concentration to extirpate nontarget amplification, which is caused by nonspecific primer dimers amplification. After 30 min of RT-LAMP reaction, the resultant amplicons were simply and rapidly analyzed by the LFA test in less than 5 min. The optimized RT-LAMP combined with the LFA allowed specific ZIKV RNA detection down to the single copy level within 35 min.

Original languageEnglish
Pages (from-to)12272-12278
Number of pages7
JournalAnalytical Chemistry
Volume88
Issue number24
DOIs
Publication statusPublished - 2016 Jan 1

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ASJC Scopus subject areas

  • Analytical Chemistry

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